Environmental Monitoring and Assessment

, Volume 105, Issue 1–3, pp 309–325

The Use of Mytilus Galloprovincialis Acetylcholinesterase and Glutathione S-Transferases Activities as Biomarkers of Environmental Contamination Along the Northwest Portuguese Coast



With the aim of using Mytilus galloprovincialis acetylcholinesterase (AChE) and glutathione S-transferases (GST) activities as biomarkers of environmental contamination in risk assessment studies along the northwest Portuguese coast, the objective of this study was to provide background information related to: (1) baseline values of these enzymatic activities both in reference and contaminated areas and their responsiveness as indicators of exposure to environmental contaminants; (2) the importance of seasonal variations of such biomarkers in M. galloprovincialis along this area. For this, the activity of these enzymes was seasonally determined in mussels collected from four sites, including a reference and three stations with different contamination sources along the referred area. Statistically significant differences on both enzymatic activities were found among the four sampling stations, at the four sampling periods. In comparison to the reference station, lower AChE and higher GST activity values were found in mussels collected in stations potentially contaminated by pesticides and domestic/industrial effluents and in mussels collected in the vicinity of an oil refinery and an industrial/mercantile harbour, respectively. The results obtained in this work highlighted the potential suitability of these biomarkers to be used as components of environmental monitoring programs in risk assessment studies along the northwest Portuguese coast. Since a seasonal variation in both enzymatic activities was found, the possible implications of such variability in the use of these enzymes as environmental biomarkers are also discussed.


acetylcholinesterase biomarkers biomonitoring coastal zone pollution , glutathione S-transferases mussels Mytilus galloprovincialis NW Portuguese coast seasonal variation 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Akcha, F., Izuel, C., Venier, P., Budzinski, H., Burgeot, T. and Narbonne, J.-F.: 2000, ‘Enzymatic biomarker measurement and study of DNA adduct formation in benzo[a]pyrene-contaminated mussels’, Mytilus galloprovincialis’, Aquat. Toxicol. 49, 269–287.Google Scholar
  2. Armknecht, S. L., Kaattari, S. L. and Van Veld, P. A.: 1998, ‘An elevated glutathione S-transferase in creosate-resistant mummichog (Fundulus heteroclitus)’, Aquat. Toxicol. 41, 1–16.Google Scholar
  3. Azevedo, D. A., Lacorte, S., Vinhas, T., Viana, P. and Barceló, D.: 2000, ‘Monitoring of priority pesticides and other organic pollutants in river water from Portugal by gas chromatography–-mass spectrometry and liquid chromatography–-atmospheric pressure chemical ionization mass spectrometry’, J. Chromatogr. A 879, 13–26.PubMedGoogle Scholar
  4. Beliaeff, B., O’Connor, T. P., Daskalakis, D. K. and Smith, P. J.: 1997, ‘U.S. mussel watch data from 1986 to 1994: Temporal trend detection at large spatial scales’, Environ. Sci. Technol. 31, 1411–1415.Google Scholar
  5. Bettencourt, A. and Ramos, L.: 2003, Estuários Portugueses, Edited by Instituto da Água, Lisbon, Portugal.Google Scholar
  6. Bradford, M.: 1976, ‘A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye binding’, Anal. Biochem. 72, 248–254.PubMedGoogle Scholar
  7. Canesi, L., Viarengo, A., Leonzio, C., Filippelli, M. and Gallo, G.: 1999, ‘Heavy metals and glutathione metabolism in mussel tissues’, Aquat. Toxicol. 46, 67–76.Google Scholar
  8. Castro, M., Santos, M. M., Monteiro, N. M. and Vieira, N.: 2001, ‘Measuring lysosomal stability as an effective tool for marine coastal environmental monitoring’, Mar. Environ. Chem. 38, 742–745.Google Scholar
  9. Clark, A. G.: 1989, ‘The comparative enzymology of the glutathione S-transferases from non-vertebrate organisms’, Comp. Biochem. Physiol. B: Biochem. Mol. Biol. 92, 419–446.Google Scholar
  10. Dellali, M., Barelli, M. G., Romeo, M. and Aissa, P.: 2001, ‘The use of acetylcholinesterase activity in Ruditapes decussatus and Mytilus galloprovincialis in the biomonitoring of Bizerta lagoon’, Comp. Biochem. Physiol. C: Pharmacol. Toxicol. 130, 227–235.Google Scholar
  11. Dias, J. M. A., Gonzalez, R., Garcia, C. and Diaz-del-Rio, V.: 2002, ‘Sediment distribution patterns on the Galicia-Minho continental shelf’, Prog. Oceanogr. 52, 249–259.Google Scholar
  12. Ellman, G. L., Courtney, D. K., Andres, V. and Featherstone, R. M.: 1961, ‘A new and rapid colorimetric determination of acetylcholinesterase activity’, Biochem. Pharmacol. 7, 88–95.PubMedGoogle Scholar
  13. Escartín, E. and Porte, C.: 1997, ‘The use of cholinesterase and carboxylesterase activities from Mytilus galloprovincialis in pollution monitoring’, Environ. Toxicol. Chem. 16, 2090–2095.Google Scholar
  14. Fitzpatrick, P. J., O’Halloran, J., Sheehan, D. and Walsh, A. R.: 1997, ‘Assessment of a glutathione S-transferase and related proteins in the gill and digestive gland of Mytilus edulis (L.), as potential organic pollution biomarkers’, Biomarkers 2, 51–56.Google Scholar
  15. Fitzpatrick, P. J. and Sheehan, D.: 1993, ‘Separation of multiple forms of glutathione S-transferase from the blue mussel’, Mytilus edulis’, Xenobiotica 23, 851–861.Google Scholar
  16. Fitzpatrick, P. J., Sheehan, D. and Livingstone, D. R.: 1995, ‘Studies on isoenzymes of glutathione S-transferase in the digestive gland of Mytilus galloprovincialis with exposure to pollution’, Mar. Environ. Res. 39, 241–244.Google Scholar
  17. Gowland, B. T. G., McIntosh, A. D., Davies, I. M., Moffat, C. F. and Webster, L.: 2002, ‘Implications from a field study regarding the relationship between polycyclic aromatic hydrocarbons and glutathione S-transferase activity in mussels’, Mar. Environ. Res. 54, 231–235.PubMedGoogle Scholar
  18. Guerreiro, N. and Pereira, P. B.: 2002, Poluição e qualidade da água, Edited by Instituto da Água, Lisbon, Portugal.Google Scholar
  19. Guilhermino, L., Barros, P., Silva, M. C. and Soares, A. M. V. M.: 1998, ‘Should the use of cholinesterases as a specific biomarker for organophosphate and carbamate pesticides be questioned?’ Biomarkers 3, 157–163.Google Scholar
  20. Habig, W. H., Pabst, M. J. and Jakoby, W. B.: 1974, ‘Glutathione S-transferases–-the first enzymatic step in mercapturic acid formation’, J. Biol. Chem. 249, 7130–7139.PubMedGoogle Scholar
  21. Hoarau, P., Gnassia-Barelli, M., Roméo, M. and Girard, J.-P.: 2001, ‘Differential induction of glutathione S-transferases in the clam Ruditapes decussatus exposed to organic compounds’, Environ. Toxicol. Chem. 20, 523–529.PubMedGoogle Scholar
  22. Kaaya, A., Najimi, S., Ribera, D., Narbonne, J. F. and Moukrim, A.: 1999, ‘Characterization of glutathione S-transferases (GST) activities in Perna perna and Mytilus galloprovincialis used as a biomarker of pollution in the Agadir Marine Bay (South of Morocco)’, Bull. Environ. Contam. Toxicol. 62, 623–629.PubMedGoogle Scholar
  23. Lacorte, S., Viana, P., Guillamon, M., Tauler, R., Vinhas, T. and Barcelò, D.: 2001, ‘Main findings and conclusions of the implementation of Directive 76/464/CEE concerning the monitoring of organic pollutants in surface waters (Portugal, April 1999–May 2000)’, J. Environ. Monit. 3, 475–482.PubMedGoogle Scholar
  24. Leal, M. C. F., Vasconcelos, M. T., Sousa-Pinto, I. and Cabral, J. P. S.: 1997, ‘Biomonitoring with benthic macroalgae and direct assay of heavy metals in seawater of the Oporto Coast (Northwest Portugal)’, Mar. Poll. Bull. 34, 1006–1015.Google Scholar
  25. Martinez-Tabche, L., Mora, B. R., Faz, C. G., Castelan, I. G., Ortiz, M. M., Gonzalez, V. U. and Flores, M. O.: 1997, ‘Toxic effect of sodium dodecylbenzenesulfonate, lead, petroleum, and their mixtures on the activity of acetylcholinesterase of Moina macrocopa in vitro’, Environ. Toxicol. Water Qual. 12, 211–215.Google Scholar
  26. Moreira, S. M., Moreira-Santos, M., Ribeiro, R. and Guilhermino, L.: 2004, ‘The Coral Bulker fuel oil spill on the North coast of Portugal: Spatial and temporal biomarker responses in Mytilus galloprovincialis’, Ecotoxicology (in press).Google Scholar
  27. Mucha, A. P., Vasconcelos, M. T. S. D. and Bordalo, A. A.: 2003, ‘Macrobenthic community in the Douro estuary: Relations with trace metals and natural sediment characteristics’, Environ. Pollut. 121, 169–180.PubMedGoogle Scholar
  28. Múrias dos Santos, A. E. F. A.: 1994, ‘Estudo e caracterização dos povoamentos bentónicos intertidais (substrato rochoso) do Norte de Portugal’, Master Thesis, Faculty of Sciences, University of Porto, Porto, Portugal.Google Scholar
  29. Narbonne, J. F., Daubèze, M., Clérandeau, C. and Garrigues, P.: 1999, ‘Scale of classification based on biochemical biomarkers in mussels: Application to pollution monitoring in European coasts’, Biomarkers 4, 415–424.Google Scholar
  30. Paris-Palacios, S., Biagianti-Risbourg, S. and Vernet, G.: 2000, ‘Biochemical and (ultra) structural hepatic perturbations of Brachydanio rerio (Teleostei, Cyprinidae) exposed to two sublethal concentrations of copper sulphate’, Aquat. Toxicol. 50, 109–124.PubMedGoogle Scholar
  31. Peakall, D. B.: 1992, Animal Biomarkers as Pollution Indicators, Chapman and Hall, London.Google Scholar
  32. Porte, C., Biosca, X., Solé, M. and Albaigés, J.: 2000, ‘The Aegean Sea oil spill on the Galician Coast (NW Spain). III: The assessment of long-term sublethal effects on mussels’, Biomarkers 5, 436–446.Google Scholar
  33. Porte, C., Solé, M., Borghi, V., Martinez, M., Chamorro, J., Torreblanca, A., Ortiz, M., Orbea, A., Soto, M. and Cajaraville, M.P.: 2001, ‘Chemical, biochemical and cellular responses in the digestive gland of the mussel Mytilus galloprovincialis from the Spanish Mediterranean coast’, Biomarkers 6, 335–350.CrossRefGoogle Scholar
  34. Power, A. and Sheehan, D.: 1996, ‘Seasonal variation in the antioxidant defense systems of gill and digestive gland of the blue mussel, Mytilus edulis’, Comp. Biochem. Physiol. C: Pharmacol. Toxicol. 114, 99–103.Google Scholar
  35. Radenac, G., Bocquené, G., Fichet, D. and Miramand, P.: 1998, ‘Contamination of a dredged-material disposal site (La Rochelle Bay, France). The use of the acetylcholinesterase activity of Mytilus edulis (L.) as a biomarker of pesticides: The need for a critical approach’, Biomarkers 3, 305–315.Google Scholar
  36. Regoli, F. and Principato, G.: 1995, ‘Glutathione, glutathione-dependent and antioxidant enzymes in mussel, Mytilus galloprovincialis, exposed to metals under field and laboratory conditions: Implications for the use of biochemical biomarkers’, Aquat. Toxicol. 31, 143–164.Google Scholar
  37. Salgado, M. A. and Serra, L.: 2001, ‘Sediment and Mussel Contamination in the Surroundings of an Oil Refinery Plant’, in: Proceedings of First International Congress on Petroleum Contaminated Soils, Sediments, and Water Analysis, Assessment and Remediation, August 14–27, 2001, Imperial College, London, U.K.Google Scholar
  38. Serra, L.: 1998, ‘Contaminação da faixa costeira de Leça da Palmeira-Parafita (Matosinhos) por hidrocarbonetos petrolíferos’, Master Thesis, Instituto de Ciências Biomédicas de Abel Salazar, University of Porto, Porto, Portugal.Google Scholar
  39. Sheehan, D. and Power, A.: 1999, ‘Effects of seasonality on xenobiotic and antioxidant defense mechanisms of bivalve molluscs’, Comp. Biochem. Physiol. C: Pharmacol. Toxicol. 123, 193–199.Google Scholar
  40. Tauler, R., Azevedo, D. A., Céspedes, R., Viana, P. and Barceló, D.: 2001, ‘Organic pollutants in surface waters from Portugal using chemometric interpretation’, Environ. Technol. 22, 1043–1054.PubMedGoogle Scholar
  41. Tauler, R., Lacorte, S., Guillamón, M., Céspedes, R., Viana, P. and Barceló, D.: 2004, ‘Chemometric modelling of main contamination sources in surface waters of Portugal’, Environ. Toxicol. Chem. 23, 565–575.PubMedGoogle Scholar
  42. Vidal, M.-L., Bassères, A. and Narbonne, J.-F.: 2002, ‘Seasonal variations of pollution biomarkers in two populations of Corbicula fluminea (Müller)’, Comp. Biochem. Physiol. C: Pharmacol. Toxicol. 131, 133–151.Google Scholar
  43. Willett, K. L., McDonald, S. J., Steinberg, M. A. and Beatty, K. B.: 1997, ‘Biomarker sensitivity for polynuclear aromatic hydrocarbon contamination in two marine fish species collected in Galveston Bay, Texas’, Environ. Toxicol. Chem. 16, 1472–1479.Google Scholar
  44. Zar, J. H.: 1996, Biostatistical Analysis, Prentice Hall International Editions, New Jersey.Google Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  1. 1.ICBAS - Instituto de Ciências Biomédicas de Abel Salazar, Departamento de Estudos de Populações, Laboratório de EcotoxicologiaUniversidade do PortoPortoPortugal
  2. 2.CIMAR - Centro Interdisciplinar de Investigação Marinha e AmbientalGrupo de EcotoxicologiaPortoPortugal

Personalised recommendations